LOCUS       sequence-485480-        7061 bp    DNA     circular SYN 08-SEP-2025
DEFINITION  Proximity-triggered Protein Trans-Splicing to generate the splice 
            product MLL-AF9, Extended Figure 4 in Nature Chemical Biology 20.10 
            (2024): 1353-1360..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7061)
  AUTHORS   Lee G, Muir TW
  TITLE     Distinct phases of cellular signaling revealed by time-resolved 
            protein synthesis.
  JOURNAL   bioRxiv [Preprint]. 2023 Jul 10:2023.07.10.548208. doi: 
            10.1101/2023.07.10.548208.
  PUBMED    37503273
REFERENCE   2  (bases 1 to 7061)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 8, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2023 Jul 10:2023.07.10.548208. doi: 
            10.1101/2023.07.10.548208."
COMMENT     Sequence Label: pCMV5-NUP98n-CAGEn-NES (Npu)
FEATURES             Location/Qualifiers
     source          1..7061
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        178..557
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        558..761
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     712..732
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     833..849
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     833..849
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             2582..2602
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2603..2623
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2843..3163
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    3262..3884
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3461..3480)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3908..3928)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3913..4242
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4093..4228
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4280..4299)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4281..4299)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4313..4329)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4313..4329)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4326..4348)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4337..4353
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4361..4367,4368..4385,4386..4391))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4406..4427
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4544..4561)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4715..5303)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4795..4814)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5474..6265,6266..6334))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6097..6116
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6335..6439)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6466..6921
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6553..6572)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6763..6784
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     join(7047..7061,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(7061,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgcc aagctgatct atacattgaa tcaatattgg caattagcca
       61 tattagtcat tggttatata gcataaatca atattggcta ttggccattg catacgttgt
      121 atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg ccatgttgac
      181 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      241 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      301 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      361 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      421 tgtatcatat gccaagtccg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      481 attatgccca gtacatgacc ttacgggact ttcctacttg gcagtacatc tacgtattag
      541 tcatcgctat taccatggtg atgcggtttt ggcagtacac caatgggcgt ggatagcggt
      601 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      661 accaaaatca acgggacttt ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg
      721 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga
      781 attcagatct ggtaccacgc gtatcgataa gcttgcatgc ctgcaggtcg actctagaac
      841 tagtggatcc gttgtgatgt ttaacaaatc atttggaaca ccctttgggg gtggcacagg
      901 tggctttggc acaacttcaa catttggaca gaatactggc tttggcacta ctagtggagg
      961 ggcatttgga acatctgcat ttggttctag caacaatact ggaggcctct ttggaaattc
     1021 acagactaaa ccaggaggat tgtttggaac cagttcattt agccagccag ctacctccac
     1081 aagcactggc tttgggtttg gtacgtcaac aggaacagca aataccttgt ttggaactgc
     1141 aagcacaggg accagtctct tctcatccca aaacaatgcc tttgcacaaa ataaaccaac
     1201 tggctttggc aattttggaa ccagtactag cagtggagga ctctttggaa ccacaaatac
     1261 cacctctaat ccttttggca gcacatctgg ctccctcttt gggccaagta gttttacagc
     1321 tgctcctact gggactacta ttaaatttaa ccctccaact ggtacagata ctatggtcaa
     1381 agctggagtt agcactaaca taagtaccaa gcaccagtgt attactgcta tgaaagaata
     1441 tgaaagcaag tcactagagg aacttcgttt agaggattat caggctaaca ggaagggccc
     1501 acagaaccag gtgggagcag gtaccacaac tggcttgttt gggtcttctc cagccacttc
     1561 cagcgcaaca ggactcttca gctcctccac cactaattca ggctttgcat atggtcagaa
     1621 caaaactgcc tttggaacta gtacaactgg atttggaaca aatccaggtg gtctctttgg
     1681 ccaacagaat cagcagacta ccagcctctt cagcaaacca tttggccagg ctacaaccac
     1741 ccagaacact ggcttttcct ttggtaatac cagcaccata ggacagccaa gcaccaacac
     1801 catgggatta tttggagtaa cccaagcctc acagcctgga ggtctttttg ggacagctac
     1861 aaacaccagc actgggacag catttggaac aggaacaggt ctctttgggc agaccaatac
     1921 tggatttggt gctgttggtt cgaccctgtt tggcaataac aagcttacta catttggaag
     1981 cagcacaacc agtgcacctt catttggtac aaccagtggc gggctctttg gttttggcac
     2041 aaataccagt gggaatagta tttttggaag taaaccagca cctgggactc ttggaactgg
     2101 gcttggtgca ggatttggaa cagctcttgg tgctggacag gcatctttgt ttgggaacaa
     2161 ccaacctaag attggagggc ctcttggtac aggagccttt ggggcccctg gatttaatac
     2221 tacgacagcc actttgggct ttggagcccc ccaggcccca gtatgtttaa gctatgaaac
     2281 ggaaatattg acagtagaat atggattatt accgattggt aaaattgtag aaaagcgcat
     2341 cgaatgtact gtttatagcg ttgataataa tggaaatatt tatacacaac ctgtagcaca
     2401 atggcacgat cgcggaaagc aaaaggtgtt tgagtattgt ttggaggatg gttcattgat
     2461 tcgggcaaca aaagaccata agtttatgac tgttgatgga caaatgttgc caattaagga
     2521 aatatttcgt cgtaagttgg atttgatgcg ggttgataat ttgccgaatg gaagcggagg
     2581 agaaaatcta tacttccaag gtgagaacct gtattttcag ggcgggagtg gtgggatcga
     2641 aatagccaca gaaaaatatt taggcgaaca aaatgtctat gacattggag ttgagcgcga
     2701 ccataatttt gcactcaaaa atggcggcta ttttcagggc atcgaaatag ccacagaaaa
     2761 atatttaggc gaacaaaatg tctatgacat tggagttgag cgcgaccata attttgcact
     2821 caaaaatggc aacagcgccg gcggagtgca ggtggaaacc atctccccag gagacgggcg
     2881 caccttcccc aagcgcggcc agacctgcgt ggtgcactac accgggatgc ttgaagatgg
     2941 aaagaaattt gattcctccc gggacagaaa caagcccttt aagtttatgc taggcaagca
     3001 ggaggtgatc cgaggctggg aagaaggggt tgcccagatg agtgtgggtc agagagccaa
     3061 actgactata tctccagatt atgcctatgg tgccactggg cacccaggca tcatcccacc
     3121 acatgccact ctcgtcttcg atgtggagct tctaaaactg gaagaacaga aactgattag
     3181 tgaggaagat ggaggaagtg gaggtagtgc tctccaaaaa aagttggaag agcttgaact
     3241 ggacgaacag tagaggatcc cgggtggcat ccctgtgacc cctccccagt gcctctcctg
     3301 gccctggaag ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc
     3361 attttgtctg actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc
     3421 aaggggcaag ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg
     3481 agtgcagtgg cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc
     3541 ctgcctcagc ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt
     3601 ttgttttttt ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa
     3661 tctcaggtga tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact
     3721 gctcccttcc ctgtccttct gattttaaaa taactatacc agcaggagga cgtccagaca
     3781 cagcataggc tacctggcca tgcccaaccg gtgggacatt tgagttgctt gcttggcact
     3841 gtcctctcat gcgttgggtc cactcagtag atgcctgttg aattgggtac gcggccagct
     3901 tggctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3961 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4021 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4081 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4141 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     4201 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctcct cgaggaactg
     4261 aaaaaccaga aagttaattc cctatagtga gtcgtattaa attcgtaatc atgtcatagc
     4321 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     4381 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     4441 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     4501 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     4561 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4621 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4681 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4741 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4801 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4861 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4921 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4981 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5041 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5101 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5161 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5221 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5281 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5341 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5401 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5461 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5521 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     5581 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     5641 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5701 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5761 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     5821 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5881 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     5941 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6001 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6061 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6121 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6181 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6241 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6301 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     6361 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6421 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg
     6481 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     6541 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     6601 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     6661 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     6721 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     6781 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     6841 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     6901 aaatattaac gcttacaatt tccattcgcc attcaggctg cgcaactgtt gggaagggcg
     6961 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     7021 attaagttgg gtaacgccag ggttttccca gtcacgacgt t
//
